Objective Parkinson's disease( PD) is a common chronic neurodegenerative disease,with four major symptoms of resting tremor, muscle rigidity, slow motion and postural balance disorder?The pathogenesis of Parkinson's disease is still unknown?A large number of studies suggested that may be the result of the interaction of genetic factors,environmental factors,aging,immune factors,specifically involved in oxidative stress,mitochondrial damage,and other mechanisms?There were 50% patients characterized by tremors,tremor is the most difficult symptoms to treat of PD, but the mechanism is still under controversial, so it ’ s of great significance to understand the generation of PD tremor, which helps to promote the clinical treatment and diagnosis.

Objective To assess the therapeutic effect of plum-blossom needle tapping combined with topical imiquimod immunotherapy on cutaneous squamous cell carcinoma (SCC) in SKH-1 mice,and to explore the immunological mechanism.Methods A total of 40 SKH-1 mice with ultraviolet light-induced cutaneous SCC were randomly and equally divided into 4 groups:control group receiving no treatment,plum-blossom needle group receiving plum-blossom needle tapping on all the tumors once a day,imiquimod group topically treated with imiquimod 5％ cream at a dose of 1.2 g/kg once a day,combination group firstly treated with plum-blossom needle tapping on all the tumors,and after the stop of bleeding topically treated with imiquimod 5％ cream at the same dose as the imiquimod group once a day.All the mice were treated for 30 days.Morphological changes of tumors in all groups were photographed and recorded every day.The tumor size was measured once every three days,and changes of total tumor volume and survival rate of the mice were compared among the 4 groups.At the end of treatment,tunor tissues were resected,and histopathological changes were compared among the 4 groups.Real-time fluorescence-based quantitative PCR (qRT-PCR) was performed to measure the mRNA expression of interferon-α (IFN-α),IFN-β,interleukin-1β (IL-1β),tumor necrosis factor-α (TNF-o) and IL-12 in tumor tissues.Results In the combination group,tumors on the back of mice grew slowly,and some even regressed.However,tumors grew fast in the control group,plum-blossom needle group and imiquimod group,and grew more slowly in the plum-blossom needle group and imiquimod group than in the control group.Before the treatment,there was no significant difference in the total tumor volume among the 4 groups (F =0.90,P ＞ 0.05).After 24-day treatment,the total tumor volume significantly differed among the 4 groups (F =5.16,P ＜ 0.05).The LSD-t test showed that the total tumor volume significantly decreased in the combination group compared with the control group (P ＜ 0.01),but no significant difference was observed among the other groups (P ＞ 0.05).Log-rank test revealed that survival curves significantly differed among the 4 groups (x2 =8.32,P ＜ 0.05).The survival rate was significantly higher in the combination group than in the control group (x2 =4.62,P =0.03),but did not differ between the plum-blossom needle group or imiquimod group and the control group or combination group (all P ＞ 0.05).Histopathological examination showed atypical cells arranged closely,a large number of tumor cells and some keratin pearls in the control group and plumblossom needle group,few dead tumor cells in the imiquimod group,and plenty of dead tumor cells,mild nuclear atypia and increased keratinization in the combination group.qRT-PCR revealed that the relative mRNA expression levels of IFN-α,IFN-β,IL-12,IL-1β and TNF-α were significantly higher in the combination group than those in the control group,plum-blossom needle group and imiquimod group (P ＜ 0.05).The imiquimod group showed significantly higher mRNA expression of IL-1β than the control group (P ＜ 0.01),but no significant differences were observed among the other groups (P ＞ 0.05).Conclusion Plum-blossom needle tapping can effectively enhance the anti-SCC activity and immunological effects of imiquimod in SKH-1 mice.

OBJECTIVE: To explore and evaluate the characteristics of spectra wave shapes and parameters of the superior vena cava (SVC) Doppler flow patterns in patients with pulmonary arterial hypertension(PAH) by echocardiography. METHODS: Forty-nine patients with PAH as the PAH group and the paired healthy subjects as the control group were included in this study. Transthoracic echocardiography was used, and the spectra of SVC flow Doppler velocities in the right supraclavicular fossa view were examined and recorded. Peak velocity and velocity time integral (VTI) were measured. RESULTS: The spectra wave shapes of SVC flow velocity in the PAH group were distinguished from those of the control. The peak velocity and VTI of S waves during inspiration and the rate of respiratory changes of peak velocity of SVC flow Doppler velocity in moderate and severe PAH groups were significantly lower than those of the control (P<0.05).The peak velocity and VTI of D waves of SVC flow Doppler velocity in the PAH groups were significantly lower than those of the control (P<0.05). The rate of respiratory changes of peak velocity of D waves in the PAH groups was significantly lower than those of the control (P<0.05). The peak velocity and VTI of AR, and VR waves of SVC flow Doppler velocity in the PAH groups were significantly higher than those of the control (P<0.05). Linear regression analysis revealed a significant correlation between pulmonary arterial systolic pressure(PASP) and the Vexp/Vins ratio (r=0.760, P<0.001). CONCLUSION The spectra wave shapes of SVC flow velocity in the PAH groups were distinguished from those of the control which may have some diagnostic value for PAH.The peak velocities and VTI of S waves during inspiration, D waves, AR waves and VR waves of SVC flow Doppler in the PAH groups are different from the control. The ratio of Vexp/Vins of S wave may be a non-invasive quantitative index to detect PAH.
Adolescent ; Adult ; Blood Flow Velocity ; Case-Control Studies ; Echocardiography, Doppler, Color ; methods ; Female ; Humans ; Hypertension, Pulmonary ; diagnostic imaging ; physiopathology ; Male ; Middle Aged ; Vena Cava, Superior ; diagnostic imaging ; physiopathology ; Young Adult

BACKGROUND: Previous studies have demonstrated that the Chinese porous tantalum made in China has non-toxicity and good biocompatibility, which can promote osteogenesis. OBJECTIVE: To investigate the effects of transforming growth factor β1 on proliferation, cel cycle and secretion of osteoblasts on porous tantalum/MG63 osteoblast-like cel composites. METHODS: Passage 3 MG63 osteoblast-like cel suspension (1×109/L) was seeded onto the porous tantalum, then the cel composites were inoculated in the medium with 0, 0.5, 5 and 10 μg/L transforming growth factor β1, respectively. The proliferation of osteoblasts was detected by cel counting kit-8 assay at 1-13 days after inoculation; the cel morphology and ultrastructure observed by scanning electron microscope and transmission electron microscopy; and level of col agen type I detected by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSON: 0.5, 5, 10 μg/L transforming growth factor β1 could promote the osteoblast proliferation, and cel proliferation in the 5 μg/L transforming growth factor β1 group was higher than that in the other groups; in the 5 μg/L transforming growth factor β1 group, laminated osteoblasts adhered on the surface and grew into inner of porous tantalum, which extended more pseudopodia toward the scaffold; osteoblasts-secreted matrix could cover the scaffold and numerous rough endoplasmic reticulum, free ribosomes, dense mitochondria, Golgi apparatus as wel as matrix vesicles could be found in the cytoplasm. In addition, the level of col agen type I in the 5 μg/L transforming growth factor β1 group was significantly higher than that in the other groups (P < 0.05). These results indicate that transforming growth factor β1 can promote proliferation, and col agen type I secretion of osteoblasts on porous tantalum/MG63 osteoblast-like cel composites, and the optimum mass concentration of transforming growth factor β1 is 5 μg/L.

Objective To understand the internal circuit contamination of ventilator in mechanical ventilation patients, to evaluate the effect of ventilator internal circuit disinfection and the impact on the incidence of ventilator-associated pneumonia (VAP). Methods A total of 39 patients with mechanical ventilation admitted to intensive care unit (ICU) of Hangzhou Geriatric Hospital from January 2017 to June 2018 were enrolled. Routine mechanical ventilation treatments for patients included pipeline replacement, aseptic operation, prevention of infection, etc. After 2 weeks of mechanical ventilation, the internal circuit of the ventilator was disinfected using the internal circuit sterilizer of the ventilator. Microorganism sampling and detection at 3 cm to the exhalation port of the internal circuit of the ventilator was performed before and after disinfection. The number of colonies was < 5 cfu/cm2 and no pathogenic bacteria could be detected. During the observation period, if the patient was complicated by VAP for anti-infective treatment, the ventilator with internal loop disinfection was replaced after infection control, and was incorporated again into the group for observation. The number of microbial colonies in the internal circuit of the ventilator before and after disinfection, the microbiological test pass rate and the incidences of VAP during the 2 weeks were observed. Results All 39 patients were included in the analysis, with 23 male and 16 female; with age of 65-97 years old, average (78.7±7.6) years old. Before the disinfection, 9 604 strains were detected in the internal circuit of the ventilator, including 8 687 strains of Gram-negative bacilli (90.4%), 902 strains of Gram-positive cocci (9.4%), and 15 strains of fungi (0.2%), which were detected in the lower respiratory tract of the patients. The strain concordance rate was 41%. The qualified rate of microbial detection in the internal circuit of the ventilator was 5.1%; 13 cases (33.3%) of VAP occurred during 2 weeks of mechanical ventilation. After disinfection, 785 strains of pathogens were detected in the internal circuit of the ventilator, and the number of colonies was significantly reduced compared with that before disinfection [cfu/cm2: 0 (0, 20) vs. 150 (15, 500), P < 0.01], of which 688 strains of Gram-negative bacilli (87.7%), 92 strains of Gram-positive cocci (11.7%) and 5 strains of fungi (0.6%) were found; the qualified rate of microbial detection in the internal circuit of ventilator reached 71.8%, which was significantly higher than that before disinfection (P < 0.01); 2 weeks after mechanical ventilation the incidence of VAP decreased slightly during the period [20.5% (8/39) vs. 33.3% (13/39)], but there was no significant difference (P > 0.05). Conclusions The internal circuit of the ventilator can be used to detect the pathogen and the sputum culture of the patients on mechanical ventilation with a high consistency. The disinfection of the pathogen could significantly reduce the air pollution of the ventilator and reduce the occurrence of VAP in the patients.

BACKGROUND:Previous studies have shown that home-made porous tantalum has non-toxicity and good biocompatibility, and can promote osteogenesis. Herein, we explore the mechanisms of tantalum-bone interface osseointegration.OBJECTIVE:To observe the morphological characteristics and expressions of integrin β1 and fibronectin on the interface between porous tantalum and bone tissues after implantation into the right rabbit femur, and to evaluate the biological mechanisms of tantalum-bone interface osseointegration.METHODS: Animal models of bilateral femoral condyle defects were made in Japanese big ear rabbits. Porous tantalum rod and allogeneic bone were respectively implanted into the left (experimental group) and right (control group) femur of rabbits. The animal specimens at the bone defect region were taken and made into paraffin sections and hard tissue sections at postoperative 2, 4, 8 weeks for morphological observation of new bone at the junction between the tantalum rod and host bone under light microscope, for osteogenic observation of the tantalum-bone interface under scanning electron microscope, and for immunohistochemical detection of integrin β1 and fibronectin expression.RESULTS AND CONCLUSION:Porous tantalum was bonded closely with the host bone. The loose and thick fibrous capsule was observed in the early stage and became thinner in the late stage shown by hematoxylin-eosin staining. The new bone was visible on tantalum-bone interface. Hard tissue slicing observation showed that the new bone was seen on the porous tantalum-bone interface, blood capillaries grew into the pores at postoperative 2 weeks and the pores were full of new bone tissues at postoperative 4 and 8 weeks. Under the scanning electron microscope, the osteoblasts appeared on the tantalum surface and in the pores at the early stage, and bone maturation and lamelar bone were seen at the late stage. The immunohistochemical results showed that the expression of integrin β1 in the experimental group was significantly lower than that in the control group at postoperative 2 weeks (P < 0.05), but the expression of fibronectin had no significant difference between the two groups (P > 0.05). In addition, there was a decline trend in the expression of integrin β1 and fibronectin atpostoperative 2, 4, 8 weeks. To conclude, the porous tantalum material is beneficial to enhance adhesion of osteoblasts on the surface and inside the micro-pores. Increased expression of integrin β1 and fibronectin on the tantalum-bone interface at early stage may promote early osteogenesis, while their decreased expression at bone maturing stage can promote osseointegration and bone remodeling.

Objective:To investigate the value of programmed death-1（PD-1） expression on the T lymphocytes for the prognosis of septic patients.Methods:From September 2017 to May 2019, septic patients were included in Department of Intensive Care Unit at 6 hospitals. The PD-1 expression on T cells were measured by flow cytometry. Logistic regression was conducted to analyze independent risk factors related to death within 28 days，and receiver operating characteristic curve(ROC) was conducted to evaluate the prognostic value of PD-1 expression on T cells in septic patients.Results:A total of 64 septic patients were enrolled to this study，including 32 survivors and 32 deaths. The PD-1 expression on T cells in the death group was significantly higher than that in the surviving group ( P<0.05). Correlation analysis showed that the percentages of PD-1 +/CD3 +T cells and PD-1 +/CD8 +T cells were positively correlated with procalciton in ( r=0.313, P =0.015； r=0.375, P=0.003), logistic regression analysis showed that the percentages of PD-1 +/CD3 +，PD-1 +/CD4 +，PD-1 +/CD8 +T cells were independent risk factors for the death of sepsis patients. The percentage of PD-1 +/CD3 +T cell was 3.63%, with AUC 0.842, sensitivity to predict the mortality 96.43% and specificity 59.38%, ( P<0.000 1). The percentage of PD-1 +/CD4 +T cell was 4.65%, with AUC 0.847, sensitivity 96.43%, specificity 62.50%，( P<0.000 1). The percentage of PD-1 +/CD8 +T cell was 3.91%, with AUC 0.771, sensitivity 64.29%, specificity 81.25%，( P=0.000 3). Conclusions:The T cell PD-1 expression is an independent risk factor to predict the 28-day mortality in septic patients. Combining the proportions of PD-1 +/CD3 +, PD-1 +/CD4 +and PD-1 +/CD8 +T cells may further enhance the predictive value for death.

Objective:To investigate the nurses' attitude towards open visitation policy in intensive care unit (ICU) and its potential influence factors, and further to provide evidence for carrying out this policy in ICU.Methods:A self-designed, anonymous online questionnaire of "attitude of ICU nurses to open visiting system in ICU" was performed in 31 provinces, autonomous regions and municipalities in China from October to December 2019, using convenient sampling method approach. Only nurses working in ICU (including specialized ICU, but excluding the critical ward in general ward) and willing to participate in the survey were included. The survey included 35 items, including the general information of each participant, the attitude towards the implementation of the open visitation system in ICU and its potential influencing factors. Ordinal Logistic regression analysis was used to identify the significant influencing factors.Results:A total of 1 558 questionnaires were sent out and 1 546 effective questionnaires were retrieved, with a response rate of 99.2%. Overall, 32.2% of them agreed with the policy, 41.3% of them disagree with the policy and 26.5% of them were uncertain. The Ordinal Logistic regression analysis showed that the independent influencing factors of ICU nurses' attitude towards open visitation policy including the possibility of increasing healthcare-associated infection [disagree: β = 1.327, 95% confidence interval (95% CI) was 0.242 to 2.413, P = 0.017; uncertain: β = 0.697, 95% CI was 0.244 to 1.151, P = 0.003], the improvement of nurses' job satisfaction (disagree: β = -1.406, 95% CI was -1.750 to -1.062, P = 0.000; uncertain: β = -0.748, 95% CI was -1.030 to -0.466, P = 0.000), the information support for medical staffs from family members (disagree: β = -0.644, 95% CI was -1.048 to -0.240, P = 0.002; uncertain: β = -0.422, 95% CI was -0.721 to -0.124, P = 0.006), the feasibility that the family members can assist the nurses in the basic nursing for patients (uncertain: β = -0.465, 95% CI was -0.729 to -0.202, P = 0.001), reducing the time that a nurse spent on caring for the patients (uncertain: β = 0.349, 95% CI was 0.052 to 0.646, P = 0.021), improving early rehabilitation (disagree: β = -0.593, 95% CI was -1.166 to -0.019, P = 0.043), and getting psychological support for patients from family members (disagree: β = 1.293, 95% CI was 0.426 to 2.159, P = 0.003), family members' satisfaction (disagree: β = -0.981, 95% CI was -1.431 to -0.531, P = 0.000). Conclusion:ICU nurses in China have realized that open visitation policy has positive effect on patients' early rehabilitation, willing to live and satisfaction; meanwhile, this policy may need more improvement in many ways such as healthcare-associated infection control, disinfection and isolation, allocation of human resources and time spent treating and caring for patients.

OBJECTIVETo detect the genomic deletion mutation in the NEMO gene of a family with incontinentia pigmenti (IP; MIM 308310).

METHODSA pedigree of IP was investigated. By using long PCR, the Delta4-10 deletion in NEMO gene was tested with specific primers In2/JF3R, and Delta4-10 deletion in pseudogene DeltaNEMO was investigated with primers Rev-2/JF3R. NEMO gene of 80 normal controls was also tested.

RESULTSThe deletion of exons 4-10 in both NEMO gene and the pseudogene DeltaNEMO was detected in all the patients in the family, but was not found in the normal individuals in this IP family and 80 unrelated controls.

CONCLUSIONThe study showed that the family with IP, which showed anticipation, was caused by NEMODelta4-10 deletion in the NEMO gene. Long PCR analysis is proven to be an efficient tool for identification of NEMO rearrangements. It could provide useful information for the genetic counseling of the family involved.

Adolescent ; Child ; Electrophoresis ; Exons ; genetics ; Family ; Female ; Humans ; I-kappa B Kinase ; genetics ; Incontinentia Pigmenti ; genetics ; Infant ; Male ; Pseudogenes ; genetics ; Sequence Deletion

OBJECTIVETo detect mutations of the NF1 gene in two sporadic cases with neurofibromatosis type 1 (NF1) and explore their molecular mechanisms.

METHODSClinical data of the two patients was collected. Genomic DNA was extracted from peripheral blood samples. Specific primers were designed to exclude pseudogenes. PCR was performed to amplify all coding exons of the NF1 gene. PCR products were directly sequenced.

RESULTSTwo novel mutations of the NF1 gene (c.1019-1020delCT in exon 9 and c.7189G to A in exon 48) were respectively identified in the two patients but not among their unaffected parents or 100 healthy controls.

CONCLUSIONMutations of the NF1 gene may have predisposed to the NF1 in the two patients.